Inflammation is a protective mechanism against pathogens, but if maintained continuously, it destroys tissue structures. Aggregatibacter actinomycetemcomitans is a gram-negative, facultative anaerobic bacterium often found in severe periodontitis. A. actinomycetemcomitans invades epithelial cells and triggers inflammatory response in the immune cells. In this study, we investigated the effect of water-soluble rosehip extract on A. actinomycetemcomitansinduced inflammatory responses. A human monocytic cell line (THP-1) was differentiated to macrophages by phorbol 12-mystristate 13-acetate treatment. The cytotoxic effect of extract was determined using the 3-(4,5-dimethylthiazol-2- yl)-2,5-diphenyltetrazolium bromide assay. The effects of extract on bacterial growth were examined by measuring the optical densities using a spectrophotometer. THP-1-derived macrophages were infected A. actinomycetemcomitans after extract treatment, and culture supernatants were analyzed for cytokine production using enzyme-linked immunosorbent assay. Protein expression was measured by western blotting. Extract was not toxic to THP-1- derived macrophages. A. actinomycetemcomitans growth was inhibited by 1% extract. The extract suppressed A. actinomycetemcomitans-induced tumor necrosis factor-α, interleukin (IL)-1β, and IL-8 production. It also decreased mitogen-activated protein kinase (MAP kinase) and nuclear factor-κB (NF-κB) phosphorylation. Moreover, the extract inhibited the expression of inflammasome components, including nucleotide-binding oligomerization domain-like receptor pyrin domain-containing protein 3, Absent in Melanoma 2, and apoptosis associated speck-like protein containing a CARD. And cysteine-aspartic proteases-1 and IL-1β expression were decreased by the extract. In summary, extract suppressed A. actinomycetemcomitans growth and decreased inflammatory cytokine production by inhibiting activation of MAP kinase, NF-κB, and inflammasome signaling. Rosehip extract could be effective in the treatment of periodontal inflammation induced by A. actinomycetemcomitans infection.

목적 : 마이봄샘기능저하증(meibomian gland dysfunction, MGD)을 수반하는 염증성 건성안의 감별진단에 대한 ‘TNF-α, IL-6, MMP-9’ 검사키트의 유용성을 평가하였다. 방법 : 건성안 이외의 안질환이 없는 20~30대 중 OSDI 설문 검사에 따른 건성안 총 118안을 대상하였고, 결막낭 메니스커스로부터 소량의 눈물을 채취하여 TNF-α, IL-6 및 MMP-9 검사를 하였다. 각막염색과 결막충혈 이 모두 Grade 1 이상인 경우는 염증성 건성안으로, 마이봄샘폐쇄와 마이봄샘구멍막힘이 모두 grade 1 이상인 경우는 MGD 관련 건성안으로 평가하였다. 염증성 건성안 및 MGD와 TNF-α, IL-6, MMP-9과의 상관성은 카 이제곱검정(Chi-square test)으로 분석하였고, ‘TNF-α, IL-6, MMP-9’ 검사키트의 염증성 건성안과 MGD를 수반하는 염증성 건성안 감별능력은 ROC 커브를 이용하여 민감도, 특이도 및 AUC(Area under the curve)를 구하고 정확도를 평가하였다. 결과 : 염증성 건성안은 TNF-α와 IL-6와 유의한 상관성을 보였고(p＜.050), ‘TNF-α, IL-6, MMP-9’ 검사 키트는 MMP-9 검사키트와 80.20%의 높은 일치도를 나타냈으나(p＜.050), 염증성 건성안 감별에 대한 민감도, 특이도, 정확도는 MMP-9 검사키트보다 낮았다. MGD는 MMP-9 검사와 상관성을 보이지 않았고, TNF-α와 IL-6 검사와는 유의한 상관성을 보였으며, MGD 감별에 대한 민감도, 특이도, 정확도는 각각 85.50%, 34.70%, 0.601, 85.50%, 32.70%, 0.591로 나타났다. MGD 수반한 염증성 건성안 감별에 대한 ‘TNF-α, IL-6, MMP-9’ 검사키트의 민감도, 특이도 및 정확도는 100.00%, 34.10%, 0.670로 MMP-9 검사키트보다 더 높았다. 결론 : MGD 진단에는 TNF-α, IL-6 검사가 유용하며, ‘TNF-α, IL-6, MMP-9’ 검사키트는 MGD를 수반한 염증성 건성안 평가에 유용할 것으로 사료된다.

To determine the effect of the tumor necrosis factor-α (TNF-α) in odontoclast formation, we administrated a TNF-α inhibitor in rats with diabetes rats with periodontitis. The rats included in the study were divided into three groups: control rats without diabetes or periodontitis (the C group), rats with periodontitis and diabetes (the PD group), and rats with periodontitis and diabetes treated by infliximab, the TNF inhibitor (the PD+infliximab group). The PD and PD+ infliximab groups received intravenous administrations of streptozotocin (STZ, 50 mg/kg) to induce diabetes. After 7 days of STZ injections, the mandibular first molars were ligatured to induce periodontitis. The PD+infliximab group was intrapenitoneally administrated by infliximab (5 mg/kg). On days 3 and 20 after the ligature administration, odontoclast formation along root surfaces was evaluated by tartrate resistant acid phosphatase (TRAP) staining and cathepsin K immunohistochemistry. On day 3, the number of TRAP- and cathepsin K-positive cells increased more so in the PD group than in the C group. The PD+infliximab group showed a lower number of positive cells than the PD group. There was no difference in all the groups on day 20. On day 3, the cathepsin-K positive multinucleated and mononucleated cells were higher in the PD group than in the C group. The number of cathepsin-K positive multinucleated cells was lower in the PD+infliximab group than in the PD group. The PD group showed more cathepsin K-positive cells in the furcation and distal surfaces than the c group. The Cathepsin K-positive cells of the PD+infliximab group were lower than that of the PD group in furcation. These results suggest that TNF-α stimulates odontoclast formation in diabetes with periodontitis.

Previous studies have investigated the potential relationship between promoter polymorphism (-308, G/ A) of tumor necrosis factor (TNF)-α and various autoimmune diseases. However, results from published data were inconclusive. To verify relationship between TNF-α polymorphism (-308, G/A) and susceptibility to autoimmune diseases such as vitiligo, celiac disease, and rheumatoid arthritis, we have performed a metaanalysis with all relevant articles before October 2016. The electronic search of PubMed, google, and Embase databases was performed to identify eligible studies investigating the relationship of TNF-α polymorphism with autoimmune diseases including vitiligo, celiac disease, and rheumatoid arthritis. Genotype frequency data of TNF-α polymorphism (-308, G/A) were extracted and the meta analysis was performed by Comprehensive meta-analysis program with odds ratio (OR) and 95% confidence intervals (95% CI). Genotype models were applied with dominant and recessive models and allele model analyzed. The final analysis included 37 publication papers with a total of 6,102 autoimmune disease patients and 6,987 control subjects. In result, a statistical significant correlation between TNF-α polymorphism (-308, G/A) and susceptibility to autoimmune disease was not detected in our meta-analysis (p>0.05 in all models). Our results suggest that the TNF-α polymorphism might not be related to the development of autoimmune disease. If further results in larger studies would be accumulated in the future, this relationship would be clarified.

Mushroom is known for anti-inflammatory and anti-oxidative potential. This study provides evidence that theinhibitory effect of mushroom on the expression of pro-inflammatory cytokines in human keratinocytes, HaCaT cells. To definethe underlying mechanisms of action, tumor necrosis factorα/IFNγ-activated human keratinocytes model was used. Mushroomsignificantly inhibited the expression of cytokines in HaCaT cells. Taken together, the results demonstrate that mushroom inhibitedinflammtion, suggesting that mushroom (DW extract: Grifola frondosa Cordyceps militaris), (Ethanol extract: Ganoderma lucidum,Lentinus edodes, Cordyceps militaris, Flammulina velutipes) might be a candidate for the treatment of skin inflammation.

Aggregatibacter actinomycetemcomitans is an important pathogen in the development of localized aggressive periodontitis. Lipopolysaccharide (LPS) is a virulent factor of periodontal pathogens that contributes to alveolar bone loss and connective tissue degradation in periodontal disease. Our present study was designed to investigate the cytokine expression and signaling pathways regulated by A. actinomycetemcomitans LPS (Aa LPS). Cytokine gene expression profiling in RAW 264.7 cells was performed by microarray analyses. The cytokine mRNA and protein levels and related signaling pathways induced by Aa LPS were measured by RT-PCR, ELISA and western blotting. Microarray results showed that Aa LPS strongly induced the expression of NF-κB, NF-κB-related genes, inflammatory cytokines, TNF-α and IL-1β in RAW 264.7 cells. NF-κB inhibitor pretreatment significantly reduced the levels of TNF-α and IL-1β mRNA and protein. In addition, the Aa LPS-induced TNF-α and IL-1β expression was inhibited by p38/JNK MAP kinase inhibitor pretreatment. These results show that Aa LPS stimulates TNF-α and IL-1β expression through NF-κB and p38/JNK activation in RAW 264.7 cells, suggesting the essential role of this pathway in the pathogenesis of localized aggressive periodontitis.

생체 내 실험에서 발효 인삼꽃 추출물(FM), 발효하지 않은 인삼꽃 추출물(FD)과 대조군으로 생리식염수를 2주간 마우스 체중 ㎏ 당 100, 200 ㎎/㎏ B.W.의 농도로 마우스에 경구 투여한 후 LPS에 의해 활성화된 복강 대식세포가 분비하는 염증성 사이토카인 IL-6, TNF-α의 생성량을 측정하였다. 그 결과, IL-6는 발효 LPS로 자극한 경우, 두 가지 농도에서 모두 처리한 군에 비해 높은 증식능을 나타내었고, LPS로 자극한 결과, 특히 발효 인삼꽃 추출물 200 ㎎/㎏ B.W. 농도에서 유의적으로 낮은 IL-6 분비량을 보였다. TNF-α의 경우, 100 ㎎/㎏ B.W.와 200 ㎎/㎏ B.W. 두 농도 모두에서 LPS로 자극하지 않은 경우, 낮은 증식 효과를 보여주었고, 자극한 경우, 인삼꽃 시료를 첨가한 군이 대조군보다 낮은 TNF-α 분비량을 보였으며, FM에서 FD보다 더 TNF-α를 억제하는 효과가 큰 것을 볼 수 있었다. 이상의 결과에 따르면 FD의 사이토카인 IL-6, TNF-α 생성 효과는 200 ㎎/㎏ B.W. 농도 투여 시 효과적으로 면역 세포와 면역 기관의 주요 기능을 증진시킬 가능성이 있을 것으로 사료된다. 이러한 연구결과를 토대로 앞으로 인삼꽃 발효를 이용한 기능성 사료 개발과 더불어 산업적 측면에서 보다 긍정적인 효과를 얻을 수 있을 것으로 판단된다.

안토시아닌(Anthocyanin)은 플라보노이드계 화합물의 한 부류로 항산화, 항암 및 항궤양, 항당뇨, 중금속해독, 시력보호, 콜레스테를 저하 등의 다양한 생리활성을 가지는 것으로 보고되어 있다. 죽상경화과정은 염증성 사이토카인의 분비 또는 혈관손상으로 인한 백혈구의 부착과 이동을 통해 시작된다. 본 연구는 이러한 죽상경화의 초기과정에서 안토시아닌 혼합물 중 single compound인 delphinidin chloride (DC) 인간혈관 내피세포

Transglutaminase 2(TGase 2) expression is modulatecl by π>JF- a in various carcinoma. The role of TGase 2 ancl TNF- a expression in salivaIY gland tumors is not clear yet. Establishecl SGT cellline has been used to study the pathogenesis of salivaIY gland adenocaI‘cinoma on a cellular level in vitro. 까le pupose of this study were to examine n버NA expression of TGase 2 and TNF- ain SGT cellline comparecl to other tumor celllines, ancl to apply these results to the paùlogenesis of salivary gland tumor. After SGT, SCC-15, HN 4, and HeLa tumor celllines were culturecl under preconfluency, ancl 3 clays after postconfluency, the cells were harvested for total RNA extraction and cDNA preparation. RT-PCR for semiquantitative mRNA analysis was done. 까le obtained results were as follows. 1. TGase 2 and π>JF- amRNA expression was not induced by confluency in all the celllines 2. TGase 2 and π'JF- amRNA expression was variable but markeclly enhanced 비 SGTcellline 3. TGase 2 n버NA expression appeared to be associated with that of π>JF- ain SGT cellline From the aboving res ults, mRNA expression of TGase 2 and TNF a should play an important role in the pathogenesis of SGT cellline originated ti'om ductal cell.

DJ#3과 DJ#13은 항염증질환 소재로서 가능성을 확인하 고자 국내 전통식품 풀질인증 된장에서 장기간 숙성된장으 로 선발된 시료이다. DJ#3과 DJ#13 추출물의 세포독성을 살펴보기 위하여 혈관내피세포를 이용하여 세포의 생존율 을 살펴본 결과 DJ#3과 DJ#13 추출물 모두 100 μg/mL의 농도까지 전혀 독성을 나타내지 않았다. 또한 DJ#3과 DJ#13 추출물의 항염증 효과를 TNF-α에 의해 활성화된 혈관내피세포에서의 NO 생성, 염증관련 단백질 발현과 mRNA 유전자 발현의 변화를 통하여 확인하였다. 혈관내 피세포에 TNF-α를 처리한 결과 NO의 함량이 유의적으로 감소하였다가 DJ#3과 DJ#13 추출물(20, 50, 100 μg/mL)을 처리하였을 때 유의성은 없으나 증가하였다. 세포배양액내 VCAM-1, ICAM-1 발현을 확인한 결과 혈관내피세포에 TNF-α를 처리한 군에서 증가된 VCAM-1 발현이 DJ#3 추출 물 20, 50 μg/mL에서 유의성 있는 감소를 보였다. 또한 DJ#3 추출물은 NO 생성과 연관 있는 eNOS mRNA의 발현을 농도 의존적으로 증가하였으며, iNOS mRNA의 발현은 농 도 의존적으로 감소하였으며 이는 NO 생성 증가가 iNOS의 발현억제를 경유한 것으로 사료된다. 또한 다수의 항염증 약물들의 작용기전이 되는 COX-2의 생성억제를 살펴본 결과 DJ#3 추출물은 TNF-α에 의해 발현되는 COX-2 단백질 의 발현을 억제하였음을 확인할 수 있었다. 또한, DJ#3 추출 물은 CAMs 단백질 및 mRNA 발현율의 감소됨을 보였다, 이상의 결과로 보아, DJ#3 추출물은 혈관내피세포에서 TNF-α로 유도된 혈관염증을 감소하는 효과를 가지고 있으 며, 항염증물질의 연구에 기초 자료로 활용이 가능할 것으 로 기대된다. 또한 염증과 관련된 cytokine 및 단백질 발현 메커니즘에 대한 추가적인 연구가 필요할 것으로 판단된다.